Boele Henk-Jan, Joung Sangyun, Fil Joanne E, Mudd Austin T, Fleming Stephen A, Koekkoek Sebastiaan K E, Dilger Ryan N
Department of Neuroscience, Erasmus MC Rotterdam, Rotterdam, Netherlands.
Princeton Neuroscience Institute, Princeton, NJ, United States.
Front Behav Neurosci. 2021 Jun 29;15:690019. doi: 10.3389/fnbeh.2021.690019. eCollection 2021.
: Pigs have been an increasingly popular preclinical model in nutritional neuroscience, as their anatomy, physiology, and nutrition requirements are highly comparable to those of humans. Eyeblink conditioning is one of the most well-validated behavioral paradigms in neuroscience to study underlying mechanisms of learning and memory formation in the cerebellum. Eyeblink conditioning has been performed in many species but has never been done on young pigs. Therefore, our aim here was to develop and validate an eyeblink conditioning paradigm in young pigs. : Eighteen intact male pigs were artificially reared from postnatal day 2-30. The eyeblink conditioning setup consisted of a sound-damping box with a hammock that pigs were placed in, which allowed the pig to remain comfortable yet maintain a typical range of head motion. In a delay conditioning paradigm, the conditional stimulus (CS) was a 550 ms blue light-emitting diode (LED), the unconditional stimulus (US) was a 50 ms eye air-puff, the CS-US interval was 500 ms. Starting at postnatal day 14, pigs were habituated for 5 days to the eyeblink conditioning setup, followed by 5 daily sessions of acquisition training (40 paired CS-US trials each day). : The group-averaged amplitude of conditioned eyelid responses gradually increased over the course of the 5 days of training, indicating that pigs learned to make the association between the LED light CS and the air-puff US. A similar increase was found for the conditioned response (CR) probability: the group-averaged CR probability on session 1 was about 12% and reached a CR probability of 55% on day 5. The latency to CR peak time lacked a temporal preference in the first session but clearly showed preference from the moment that animals started to show more CRs in session 2 and onwards whereby the eyelid was maximally closed exactly at the moment that the US would be delivered. : We concluded that 3-week-old pigs have the capability of performing in a cerebellar classical conditioning task, demonstrating for the first time that eyeblink conditioning in young pigs has the potential to be a valuable behavioral tool to measure neurodevelopment.
猪已成为营养神经科学中越来越受欢迎的临床前模型,因为它们的解剖结构、生理机能和营养需求与人类高度相似。眨眼条件反射是神经科学中最经过充分验证的行为范式之一,用于研究小脑学习和记忆形成的潜在机制。眨眼条件反射已在许多物种中进行,但从未在幼猪身上进行过。因此,我们这里的目的是开发并验证幼猪的眨眼条件反射范式。
18只完整的雄性猪从出生后第2天到第30天进行人工饲养。眨眼条件反射装置由一个带有吊床的隔音箱组成,猪被放置在吊床上,这使得猪既能保持舒适,又能保持典型的头部运动范围。在延迟条件反射范式中,条件刺激(CS)是一个550毫秒的蓝色发光二极管(LED),无条件刺激(US)是一个50毫秒的眼部吹气,CS-US间隔为500毫秒。从出生后第14天开始,猪对眨眼条件反射装置进行5天的习惯化训练,随后每天进行5次获取训练(每天40次配对的CS-US试验)。
在5天的训练过程中,条件性眼睑反应的组平均幅度逐渐增加,这表明猪学会了将LED光CS与吹气US联系起来。条件反应(CR)概率也有类似的增加:第1次训练时的组平均CR概率约为12%,到第5天达到了55%的CR概率。CR峰值时间的潜伏期在第一次训练中没有时间偏好,但从动物在第2次训练及以后开始表现出更多CR时起,就明显表现出偏好,即在美国即将施加时,眼睑正好最大限度地闭合。
我们得出结论,3周龄的猪有能力完成小脑经典条件反射任务,首次证明幼猪的眨眼条件反射有潜力成为测量神经发育的有价值的行为工具。